scholarly journals Deoxyribonuclease from Salmon Testes

1962 ◽  
Vol 45 (4) ◽  
pp. 77-92 ◽  
Author(s):  
Margaret R. McDonald
Keyword(s):  
Ionic Strength ◽  
Maximal Activity ◽  
Optimum Concentration ◽  
Heat Denaturation ◽  
Acid Extraction ◽  
Fractional Precipitation ◽  
Temperature Optimum ◽  
Monovalent Ions ◽  
Ph Range ◽  
Deoxyribonuclease Activity

A procedure is described for the purification of salmon testis deoxyribonuclease II by means of acid extraction, fractional precipitation with ammonium sulfate, heat denaturation of extraneous proteins, and ethanol fractionation. This process separates the deoxyribonuclease activity from that of ribonuclease, phosphatase, phosphodiesterase, and protease. Over 50 per cent of the activity is retained with an over-all enrichment of 20,000-fold. The enzyme degrades both native and heat-denatured DNA, but the rate of degradation of the latter is only one-tenth that of the former. It does not hydrolyze apurinic acid. The enzyme is most stable in the pH range 4 to 5. Electrolytes are essential for the expression of its activity: monovalent ions satisfy the requirement, but divalent ones are much more effective. Above a certain optimum concentration, each electrolyte is inhibitory. The pH of maximal activity, under conditions of optimal ionic strength, is 4.8; the temperature optimum is near to 55°C.

Biological characteristics of an enterotoxin produced by Bacillus cereus

1975 ◽  
Vol 21 (8) ◽  
pp. 1236-1246 ◽  
Author(s):  
W. M. Spira ◽  
J. M. Goepfert
Keyword(s):  
Ionic Strength ◽  
Bacillus Cereus ◽  
Exponential Growth ◽  
Biological Activities ◽  
Egg Yolk ◽  
Immune Serum ◽  
Fluid Accumulation ◽  
Ileal Loop ◽  
Receptor Sites ◽  
Ph Range

An enterotoxin synthesized during exponential growth by Bacillus cereus produces fluid accumulation in rabbit ileal loops, alters vascular permeability in the skin of rabbits, and kills mice when injected intravenously. All activities are eluted simultaneously from a Sephadex G-75 column and are distinct from the hemolysin and egg yolk turbidity factor of B. cereus. The enterotoxin is a true exotoxin. It interacts with intestinal receptor sites in a highly transient manner in the ileal loop system. Rabbit immune serum produced against the culture fluids from one strain of B. cereus neutralized the three biological activities in all other strains tested except strain B-6-ac for which none of the activities were neutralized.Enterotoxin proved to be unstable under a wide variety of conditions; ionic strength was especially critical. Enterotoxin was most stable in a pH range of 5.0 to 10.0, but lost activity rapidly outside this range. Alkylation provided some protection of enterotoxin activity in crude preparations but failed to protect activity during purification procedures. It did not appear to affect critically the enterotoxin molecule itself, since elution profiles on Sephadex G-75 chromatography were unchanged after alkylation.

Why is the hydrolytic activity of acetylcholinesterase pH dependent? Kinetic study of acetylcholine and acetylthiocholine hydrolysis catalyzed by acetylcholinesterase from electric eel

2018 ◽  
Vol 73 (9-10) ◽  
pp. 345-351 ◽  
Author(s):  
Alena Komersová ◽  
Markéta Kovářová ◽  
Karel Komers ◽  
Václav Lochař ◽  
Alexander Čegan
Keyword(s):  
Catalytic Activity ◽  
Ionic Strength ◽  
Ph Value ◽  
Hydrolytic Activity ◽  
Initial Substrate ◽  
Electric Eel ◽  
Value Range ◽  
Ph Dependent ◽  
Ph Range ◽  
Initial Substrate Concentration

AbstractThe dependence of the activity of acetylcholinesterase from electric eel at a pH value range of 4.8–9.8 (phosphate buffer), regarding acetylcholine and acetylthiocholine hydrolysis, was determined at 25 °C, ionic strength of 0.11 M, and initial substrate concentration of 4 mM. At a pH range of 4.8–9.8, the dependencesA(pH) form a sigmoid increasing curve with the maximum catalytic activity at a pH range 8–9.5. For acetylcholine hydrolysis, the kinetic reason for such an increase inAconsists mainly of an increase in the rate constantk2(Michaelis-Menten) model with increasing pH of the reaction mixture. For acetylthiocholine hydrolysis, the kinetic explication of the determined dependenceA(pH) is more complicated.

Regulatory features of transcription in isolated mitochondria from Artemia franciscana embryos

1999 ◽  
Vol 277 (6) ◽  
pp. R1588-R1597 ◽  
Author(s):  
Brian D. Eads ◽  
Steven C. Hand
Keyword(s):  
Rna Synthesis ◽  
Maximal Activity ◽  
Artemia Franciscana ◽  
Mitochondrial Rna ◽  
Optimal Conditions ◽  
In Organello ◽  
Atp Inhibition ◽  
Bona Fide ◽  
Ph Range

Optimal conditions were developed for an in organello transcriptional run-on assay using mitochondria isolated from Artemia franciscana embryos to investigate potential regulatory features of RNA synthesis under conditions of anoxia-induced quiescence. Transcription is not dependent on oxidative phosphorylation for maximal activity when exogenous ATP is available. Bona fide transcription products, as assessed by hybridization with specific mitochondrial cDNAs from A. franciscana, are produced in an inhibitor-sensitive manner. Transcription rate measured at pH 7.9 is reduced 80% when pH is lowered to 6.3, a pH range that mimics the in vivo change seen on exposure of embryos to anoxia. The proton sensitivity of mitochondrial RNA synthesis may provide a mechanism to depress this significant energy expenditure during quiescence. The influence of nucleotide concentration on kinetics is complicated by an interdependence among nucleotide species. ATP inhibition observed at subsaturating UTP concentrations is relieved when UTP is at saturating, physiologically relevant levels. Taken together, these data suggest that local (versus nuclear mediated) control is important in dictating mitochondrial transcription during rapid modulations in gene expression, such as those observed under anoxia-induced quiescence.

scholarly journals Improvement in Thermostability of an Achaetomium sp. Strain Xz8 Endopolygalacturonase via the Optimization of Charge-Charge Interactions

2015 ◽  
Vol 81 (19) ◽  
pp. 6938-6944 ◽  
Author(s):  
Tao Tu ◽  
Huiying Luo ◽  
Kun Meng ◽  
Yanli Cheng ◽  
Rui Ma ◽  
...  
Keyword(s):  
Rational Design ◽  
Residual Activity ◽  
Maximal Activity ◽  
Wild Type ◽  
Content Type ◽  
Highly Active ◽  
Wide Ph Range ◽  
Enzyme Thermal Stability ◽  
Ph Range ◽  
Charge Interactions

ABSTRACTImproving enzyme thermostability is of importance for widening the spectrum of application of enzymes. In this study, a structure-based rational design approach was used to improve the thermostability of a highly active, wide-pH-range-adaptable, and stable endopolygalacturonase (PG8fn) fromAchaetomiumsp. strain Xz8 via the optimization of charge-charge interactions. By using the enzyme thermal stability system (ETSS), two residues—D244 and D299—were inferred to be crucial contributors to thermostability. Single (D244A and D299R) and double (D244A/D299R) mutants were then generated and compared with the wild type. All mutants showed improved thermal properties, in the order D244A < D299R < D244A/D299R. In comparison with PG8fn, D244A/D299R showed the most pronounced shifts in temperature of maximum enzymatic activity (Tmax), temperature at which 50% of the maximal activity of an enzyme is retained (T50), and melting temperature (Tm), of about 10, 17, and 10.2°C upward, respectively, with the half-life (t1/2) extended by 8.4 h at 50°C and 45 min at 55°C. Another distinguishing characteristic of the D244A/D299R mutant was its catalytic activity, which was comparable to that of the wild type (23,000 ± 130 U/mg versus 28,000 ± 293 U/mg); on the other hand, it showed more residual activity (8,400 ± 83 U/mg versus 1,400 ± 57 U/mg) after the feed pelleting process (80°C and 30 min). Molecular dynamics (MD) simulation studies indicated that mutations at sites D244 and D299 lowered the overall root mean square deviation (RMSD) and consequently increased the protein rigidity. This study reveals the importance of charge-charge interactions in protein conformation and provides a viable strategy for enhancing protein stability.

Formation and Properties of Americium Colloids in Aqueous Systems

1981 ◽  
Vol 6 ◽  
Author(s):  
U. Olofsson ◽  
B. Allard ◽  
K. Andersson ◽  
B. Torstenfelt
Keyword(s):  
Ionic Strength ◽  
Aqueous Solutions ◽  
Storage Time ◽  
Large Fraction ◽  
Average Charge ◽  
Particle Sizes ◽  
Aqueous Systems ◽  
Ph Range ◽  
And Storage ◽  
Solution Changes

ABSTRACTThe formation and sorption properties of colloidal americium in aqueous solutions have been studied with variations of pH (2–12), ionic strength (0.01–1.0 M NaClO4), americium concentration (10−7 −10−11 M) and storage time (6 h–6 months). A large fraction of the americium is sorbed on the container walls or on Al2O3 in the pH-range 7–11. Around pH 5–8 and at pH above 12 centrifugable fractions (particle sizes greater than 100 nm) are obtained. The fraction formed at pH above 12, which seems to be a true hydroxide colloid, migrates through an A12O3-filled column with very little retention. The average charge of americium species in solution changes from positive values at pH below 7–8 to negative values at pH above 10–11, as indicated from electromigration studies.

scholarly journals Effect of pH, ionic strength and univalent inorganic ions on the reconstitution of aspartate aminotransferase

1974 ◽  
Vol 137 (2) ◽  
pp. 199-203 ◽  
Author(s):  
Liliana Gianfreda ◽  
Gennaro Marino ◽  
Rosaria Palescandolo ◽  
Vincenzo Scardi
Keyword(s):  
Ionic Strength ◽  
Aspartate Aminotransferase ◽  
Pyridoxal Phosphate ◽  
Water Structure ◽  
Inorganic Ions ◽  
Halide Ions ◽  
Ph Change ◽  
Effect Of Ph ◽  
Ph Range ◽  
Ph Curve

1. The effect of pH change on the reconstitution of aspartate aminotransferase (EC 2.6.1.1), i.e. the reactivation of the apoenzyme with coenzyme (pyridoxal phosphate and pyridoxamine phosphate), was studied in the pH range 4.2–8.9 by using three buffer systems at concentrations ranging from 0.025 to 0.1m. 2. Although the profile of the reconstitution rate–pH curve in the range pH5.2–6.8 (covered by sodium cacodylate–HCl buffer) reflects the influence of the H+ concentration on the reconstitution process, the profile of the curve in the pH ranges 4.2–5.6 and 7.2–8.25 (covered respectively by sodium acetate–acetic acid and Tris–HCl buffers) appears to be influenced by the ionic strength of the buffer. 3. The reconstitution is also influenced by univalent inorganic ions such as halide ions and, to a lesser extent, alkali metal ions, which are known to alter the water structure.

Kinetics and mechanism of disproportionation and ferricyanide oxidation of the 10-methylacridinium cation in aqueous base

1984 ◽  
Vol 62 (4) ◽  
pp. 729-735 ◽  
Author(s):  
John W. Bunting ◽  
Glenn M. Kauffman
Keyword(s):  
Ionic Strength ◽  
Second Order ◽  
Kinetics And Mechanism ◽  
Oxidation Pathway ◽  
First Order ◽  
Aqueous Base ◽  
Ph Range ◽  
Rate Profile ◽  
Kinetics Of ◽  
Major Oxidation

The kinetics of disproportionation and ferricyanide ion oxidation of the 10-methylacridinium cation have been measured spectrophotometrically over the pH range 9–14 in.20% CH3CN – 80% H2O (v/v) and ionic strength 1.0 at 25 °C. Disproportionation is kinetically second-order in total acridine species. The pH–rate profile is consistent with the rate-determining reaction of one acridinium cation with the pseudobase alkoxide anion derived from a second acridinium cation. Ferricyanide ion oxidation is kinetically first-order in each of ferricyanide ion and total acridine species. The pH–rate profile requires three distinct pathways for the ferricyanide ion oxidation of the 10-methylacridinium cation. For pH < 9.7, rate-determining attack of ferricyanide ion on the neutral pseudobase predominates, while for pH > 12.8 the predominant oxidation pathway involves reaction of ferricyanide ion with the pseudobase alkoxide ion. Between pH 9.7 and 12.8, the major oxidation pathway involves initial disproportionation of the acridinium cation followed by ferricyanide ion oxidation of the 9,10-dihydro-10-methylacridine product. This latter route accounts for a maximum of 69% of the total ferricyanide ion oxidation at pH 11.1.

scholarly journals Evaluation of the LIVE/DEAD BacLight Kit for Detection of Extremophilic Archaea and Visualization of Microorganisms in Environmental Hypersaline Samples

2004 ◽  
Vol 70 (11) ◽  
pp. 6884-6886 ◽  
Author(s):  
Stefan Leuko ◽  
Andrea Legat ◽  
Sergiu Fendrihan ◽  
Helga Stan-Lotter
Keyword(s):  
Ionic Strength ◽  
Potential Application ◽  
Environmental Samples ◽  
High Ionic Strength ◽  
Ph Range

ABSTRACT Extremophilic archaea were stained with the LIVE/DEAD BacLight kit under conditions of high ionic strength and over a pH range of 2.0 to 9.3. The reliability of the kit was tested with haloarchaea following permeabilization of the cells. Microorganisms in hypersaline environmental samples were detectable with the kit, which suggests its potential application to future extraterrestrial halites.

scholarly journals A DEMONSTRATION OF SEVERAL DEOXYRIBONUCLEASE ACTIVITIES IN MAMMALIAN CELL MITOCHONDRIA

1974 ◽  
Vol 62 (3) ◽  
pp. 695-706 ◽  
Author(s):  
Michael Durphy ◽  
Paul N. Manley ◽  
Errol C. Friedberg
Keyword(s):  
Ionic Strength ◽  
Adult Rabbit ◽  
Mitochondrial Enzyme ◽  
Alkaline Ph ◽  
Sh Group ◽  
Liver And Kidney ◽  
Substrate Variation ◽  
Enzyme Markers ◽  
Triton X ◽  
Deoxyribonuclease Activity

Extracts of purified mitochondria from adult rabbit liver and kidney have been prepared by lysis with Triton X-100. Such extracts contain deoxyribonuclease activity demonstrable at alkaline pH. Studies utilizing the effects of substrate variation, differing ionic strength, nucleoside di- and triphosphates, and SH-group inhibitors reveal the existence of at least five distinguishable deoxyribonuclease activities in these extracts. Assay of lysosomal and mitochondrial enzyme markers indicates no significant lysosomal contamination of the mitochondrial extracts. Further studies also suggest that the alkaline deoxyribonuclease activity is specifically located in or in association with mitochondria.

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